Population of antibodies comprising homogeneous antibodies each having left-right asymmetric sugar chains, and method for producing same

Abstract

A population of antibodies including homogeneous antibodies in which N-linked complex sugar chains attached to asparagine (Asn) at position 297 of CH domains of Fc regions of two heavy chains on the left and the right of each antibody are sugar chains structurally different from each other is described as well as a method of producing the population of antibodies.

1 . A method for fractionating antibodies in a population of antibodies into a non-glycosylated antibody fraction, an antibody glycosylated at one heavy chain fraction, and an antibody glycosylated at two heavy chains fraction, comprising contacting a population of antibodies with an affinity chromatography carrier having an Fcγ receptor immobilized thereon as a ligand and thereafter recovering the non-glycosylated antibody fraction, an antibody glycosylated at one heavy chain fraction, and an antibody glycosylated at two heavy chains fraction,
 wherein the elution time is shorter in the order of a non-glycosylated antibody fraction, an antibody glycosylated at one heavy chain fraction, and an antibody glycosylated at two heavy chains fractions. 
 
     
     
         2 . The method of  claim 1 , wherein the population of antibodies is treated with endo-β-N-acetylglucosaminidase (ENGase) prior to the population of antibodies contacting the carrier. 
     
     
         3 . The method of  claim 2 , wherein the ENGase is EndoS. 
     
     
         4 . The method of  claim 2 , wherein the ENGase is selected from the group consisting of EndoS D233Q, Endo S2 D184M, Endo S2 D184Q, EndoS D233Q/Q303L, D233Q/A303L/E350Q, and Endo M N175Q, and which is modified so as to suppress sugar chain cleavage activity and improve sugar chain transfer activity.